Open AccessPeriodontal pathogens: A quantitative comparison of anaerobic culture and real‐time PCR
Author(s) -
Boutaga Khalil,
Winkelhoff Arie Jan,
VandenbrouckeGrauls Christina M.J.E.,
Savelkoul Paul H.M.
Publication year - 2005
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1016/j.femsim.2005.03.011
Subject(s) - prevotella intermedia , actinobacillus , microbiology and biotechnology , biology , fusobacterium , treponema denticola , fusobacterium nucleatum , aggregatibacter actinomycetemcomitans , porphyromonas gingivalis , peptostreptococcus , periodontitis , real time polymerase chain reaction , prevotella , bacteroidaceae , periodontal disease , polymerase chain reaction , 16s ribosomal rna , microbiological culture , anaerobic exercise , anaerobic bacteria , bacteroides , bacteria , gene , medicine , genetics , dentistry , physiology
Periodontitis is a multi‐factorial chronic inflammatory and destructive disease of the tooth‐supporting tissues. Quantitative anaerobic culture techniques have been used for microbial diagnosis of the different forms of the disease. The aim of this study was to compare real‐time PCR with quantitative anaerobic culture for detection and quantification of 5 prominent periodontal pathogens. Real‐time PCR assays with the 16s rRNA genes of Actinobacillus actinomycetemcomitans , Prevotella intermedia , Tannerella forsythensis , Peptostreptococcus micros and Fusobacterium spp. were developed. The PCR was validated on pure cultures of various bacterial strains. Subsequently, subgingival plaque samples from 259 adult patients with periodontitis were analyzed with quantitative anaerobic culture and real‐time PCR. A standard curve for DNA quantification was created for each primer‐probe set based on colony‐forming units equivalents. All bacterial species were correctly identified. The lower limits of detection by PCR varied between 1–50 colony‐forming units equivalents depending on the species. No cross‐reactivities with heterologous DNA of other bacterial species were observed. Real‐time PCR results showed a high degree of agreement with anaerobic culture results. Real‐time PCR is a reliable alternative for diagnostic quantitative anaerobic culture of subgingival plaque samples.